UVAbsorbance(280nm)–ProteinDetermination
UV Absorbance (280 nm) – Protein Determination Simple and quick method to accurately quantitate total protein in purified material or approximately quantitate total protein in crude lysates or partial purified material.(Protein Determination by UV Absorption - Alastair Aitken and Michale Learmonth - Protein Protocols in CD Rom - Humana Press, 1998)IntroductionQuantitation of the amount of protein in a solu......閱讀全文
Protein－Electrophoresis
DefinitionAmino acids, nucleotides, polypeptides, and other compounds in a colloidal state can be separated by the application of external voltages wh
Spectrophotometry
A spectrophotometer or colorimeter makes use of the transmission of light through a solution to determine the concentration of a solute within the so
Labeling－Tubulin－and－Quantifying－Labeling－Stoichiometry2
II. Labeling ProtocolThe procedure described below can be scaled down if desired. It is essential to perform all steps involving caged dyes under a sa
Protein－Kinase－A－at－the－Centrosome
Protein kinase A regulatory subunit RIIalpha (PKA-RIIa) is tightly bound to centrosomal structures during interphase through
Acetone－precipitation－of－protein
This procedure is suitable for recovering proteins from most aqueous solvents and from SDS containing buffers. It is not recommended for proteins dis
Preparing－a－Selenomethionyl－Protein
Purpose The protocol describes how to prepare selenomethionyl (Se-Met) protein using a regular E.coli strain. Selenium can be used for phase determi
Protein－arginine－methylation
Typical modification sites: RGG box or RXR sequence motifs R-arginine, G-glycine, X-any aminoacid. Enzymes catalysing protein arginine methylation: P
Eukaryotic－protein－translation
The scanning translation initiation model suggests that 40S ribosomal subunit preloaded with factors bind to the 5’ end of the mRNA near the cap. The
Protein－A－Purification－of－Antibody
1. Reagents(1) Affi-gel Protein-A Agarose (BioRad #153-6153)(2) MAPS II Binding Buffer (BioRad # 153-6161)(3) 0.314 g/ml diH2O(4) MAPS II Elution Buff
Protein－purification;－actin
Protein purification; actin ? ? ?Overview?? ACTINThe most abundant muscle and non-muscle cytoskeletal protein. MW 42 kDa, 374/375 amino acids; various
Protein－Staining－Procedures
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor
Bradford－protein－assay
Bradford protein assayConsiderations for useThe Bradford assay is very fast and uses about the same amount of protein as the Lowry assay. It is fairly
Protein－Assay－(Spectrophotometer)
Protein Assay (Spectrophotometer)Use BSA (bovine serum albumin) 1mg/ml stock solution (1ml Eppendorf tubes) for standard curve.Place 0, 2, 5, 10, 15,
LOWRY－PROTEIN－ASSAY
The Lowry procedure is one of the most venerable and widely-used protein assays, being first described in 1951 [Lowry et al.,?J. Biol. Chem. 193: 265-
Appendix－G:－Spectrophotometry
Figure G.1 Electromagnetic Radiation SpectrumFigure G.2. Schematic light transmissionFigure G.3. Use of the Spec 20A spectrophotometer or colorimeter
UV－Shadowing
UV shadowing is a technique for visualizing nucleic acids separated on acrylamide/urea gels. The technique utilizes shortwave UV light (254 nm) and a
分子克隆蛋白表達實驗指南（十二）
– Note: The yield of fusion protein can be estimated by measuring the ? absorbance at 280 nm. The GST affinity tag can be approximated by 1 A280 ? 0.5
Basic－Protein－Chemistry－Techniques
Coomassie Blue Stain:? (for gels)? 1) Combine 225 ml Methanol with 225 ml ddH2O.? 2) Add 0.5 grams of Coomassie Blue.? 3) Just before use, add 50 ml
Mechanism－of－Protein－Import－into－the－Nucleus
Nuclear pore complexes (NPCs) are large proteinaceous assemblies that provide the only known portals for exchang
Basic－Protein－Chemistry－Techniques
實驗概要 Basic Protein Chemistry Techniques 實驗步驟 Coomassie Blue Stain:? (for gels)? 1) Combine 225 ml Methanol with 225 ml ddH2O.? 2) Add 0.5 grams of
Angiotensin－Protein－Kinase－Assay
James Hardwick's angiotensin assay protocol This specific procedure was developed to assay the activity of the Lck kinase expressed from a retroviral
Protein－G－Purification－of－Antibodies
1. Reagent and Materials(1) Hi-Trap Protein G Column (Pharmacia Biotech #17-0404-01)(2) 20 mM Sodium Phosphate Buffer, pH 7.01.084 g NaH2PO4, anhydrou
Protocol－for－Protein－Extraction－for－proteomics
Protocol for Protein Extraction10 % w/v TCA/ acetone/ 0.07 % v/v -MercaptoethanolPlant cells are rich in compounds that interfere with the 2DE separat
Protein－Immunolocalization－in－Maize－Tissues
The ?analysis of gene expression at transcript and at protein level is of ?outstanding importance in plant developmental biology. Proteins can be ?loc
Cyanogen－Bromide－digestion－of－protein
1. Proteins are TCA precipitated and washed with acetone, then dried.2. The CNBr should be brought to room temperature in the hood and used ONLY in th
Coupling－Antibodies－to－Protein－A－or－G
1. use 2 mg of antibody per ml wet beads (use appropriate antibody/protein A or G combination).2. mix antibodies with beads and bind at room temperatu
Western－Blot－with－Platelet－Protein
OUTLINEWestern blot is a wide used technique to identify a target protein/s for the certain antibody.PROTOCOLPrepare platelets.Lyse washed platelets (
In－Vitro－Protein－Ubiquitination－Assay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquiti
Biorad－Protein－Assay:－Bradford
Biorad Protein Assay: BradfordStandards: 1 mg/ml BSA stock- dilute 1:10 to get 0.1 mg/ml BSAAdd To get H-2O20 μl 2 μg/ml 780 μl40 μl 4 μg/ml 760 μl60
Protein－Expression－and－Purification－Protocol
Step 1:?Transform?appropriate DNA plasmid into BL21(DE3)?E. coli?cells. These cells must be competent. (Protocol for how to make competent cells.)a) T